This application broad Challenge Area 06 Enabling Technologies and specific Challenge Topic 06-CA- 117. Technologies that provide new tools and insights for basic research with increased speed, cost efficiency, sensitivity, selectivity, or the capability to create new avenues of research into the specific mechanisms that can lead to a better understanding of the development and progression of cancer. Of interest are technologies for molecular, subcellular, cellular and extracellular structure/function studies;capture, separation, and characterization of biomolecules, molecular complexes, sub-cellular complexes, cells, and complex mixtures;and technologies to facilitate the development of more accurate in vitro and in vivo cancer models (especially mouse models for human cancers). Of specific interest are new technologies that enhance understanding of the tumor microenvironment, cancer stem cells, complex pathways, and the role of pathogens in cancer development. The human papillomaviruses (HPVs) are associated with a variety of human cancers. Over 100 different HPVs have now been identified and about 25 of these have been associated with human cancer. The papillomaviruses can be grouped into a phylogenetic tree based on sequence relatedness. The cancer associated HPVs fall into two different groups, the genus alpha papillomaviruses and the genus beta papillomaviruses. The HPVs of genus alpha primarily infect mucosal epithelium of the genital tract and the upper airway, and the HPVs of the genus beta cause cutaneous lesions. The HPVs in each of these genera are further grouped by sequence relatedness into species that are somewhat predictive of biological behavior and cancer risk. Among the alpha-HPVs, the high-risk types (including HPV16, HPV18, HPV31, HPV33, HPV45 and HPV52) that are associated with cervical cancer are found in species 7 and 9. The low-risk HPVs, such as HPV6 and HPV44 associated with venereal warts, are in species 10. HPV2 and HPV57 are not associated with any cancers and do not cause genital mucosal lesions and are in species 4. The role of the species 7 and 9 members of the alpha-HPVs in cancer has now well been established by both molecular and epidemiologic studies. Approximately 90% of human cervical cancers harbor the DNA of a high-risk HPV type, and express two genes (E6 and E7) that function as viral oncogenes. Much of our understanding of the mechanisms by which the genus alpha HPVs contribute to cancer progression comes from the identification of the cellular proteins that bind and interact with HPV encoded proteins. The HPVs of the beta-genus cause cutaneous lesions. Many of these HPVs were first identified in skin lesions and cancers in patients with the rare dermatologic disease epidermodysplasia verruciformis (EV). These HPV types have also been found in squamous cell skin cancers (SCSC) from both immunosuppressed and immunocompetent individuals, and they have been found in skin swabs and eyebrow samples from normal individuals. The mechanistic role of the genus beta HPV types in human SCSC is less clear. The association of some of these HPVs (such as HPV5 and HPV8) with SCSC is stronger than for others. It may be that, like the genus alpha viruses, some of the genus beta viruses may present a higher risk for cancer progression than others. From a molecular standpoint, the genus beta viruses have not yet been well studied and the mechanisms by which they may contribute to cancer progression remain to be elucidated. This Challenge Grant proposal combines expertise from the Harper and Howley laboratories to provide new HPV tools, and to apply CompPASS, a new computational and informatic platform for proteomic analysis of protein complexes and interaction networks, to provide new insights into the biology of the oncogenic HPVs and the mechanisms by which they cause human cancer. PUBLIC HEALTH RELEVANCE: The human papillomaviruses are a family of viruses that are associated with a number of human cancers. Important insights into the cellular pathogenesis associated with these viruses and how they cause cancer have come from the identification of the cellular proteins that are targeted by HPV virally encoded proteins. We will generate a set of research tools for the viral proteins encoded by 18 different HPV types representing two distinct genera of HPVs that have been associated with cancer and will determine the cellular proteins and pathways with which they interact.